Advanced search
Start date
Betweenand
Related content
(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Human bone morphogenetic protein-2 (hBMP-2) characterization by physical-chemical, immunological and biological assays

Full text
Author(s):
Show less -
Suzuki, Miriam Fussae [1] ; Oliveira, Joao Ezequiel [1] ; Damiani, Renata [2] ; Lima, Eliana Rosa [1] ; Amaral, Kleicy Cavalcante [1] ; Santos, Anderson Maikon de Souza [3, 4] ; Magalhaes, Geraldo Santana [5] ; Faverani, Leonardo Perez [3, 4] ; Pereira, Luis Antonio Violin Dias [6] ; Silva, Fabiana Medeiros [2] ; Bartolini, Paolo [1]
Total Authors: 11
Affiliation:
[1] IPEN CNEN SP, Inst Pesquisas Energet & Nucl, Ctr Biotechnol, Ave Prof Lineu Prestes 2242, BR-05508000 Sao Paulo, SP - Brazil
[2] Biosintesis P&D, Sao Paulo, SP - Brazil
[3] Univ Estadual Paulista Julio de Mesquita Filho UN, Sch Dent, Dept Surg, Aracatuba, SP - Brazil
[4] Univ Estadual Paulista Julio de Mesquita Filho UN, Sch Dent, Integrated Clin, Aracatuba, SP - Brazil
[5] Inst Butantan, Immunopathol Lab, Sao Paulo, SP - Brazil
[6] Univ Estadual Campinas UNICAMP, Inst Biol, Dept Biochem & Tissue Biol, Campinas, SP - Brazil
Total Affiliations: 6
Document type: Journal article
Source: AMB EXPRESS; v. 10, n. 1 FEB 17 2020.
Web of Science Citations: 0
Abstract

Commercially available preparations of methionyl-human BMP-2 and CHO-derived hBMP-2, which belongs to the transforming growth factor beta (TGF-beta) superfamily, were used for a complete characterization. This protein is an extremely efficient osteoinductor that plays an important role during bone regeneration and embryonic development. Characterization was carried out via SDS-PAGE and Western blotting, followed by reversed-phase HPLC, size-exclusion HPLC and MALDI-TOF-MS. The classical in vitro bioassay, based on the induction of alkaline phosphatase activity in C2C12 cells, confirmed that hBMP-2 biological activity is mostly related to the dimeric form, being 4-fold higher for the CHO-derived glycosylated form when compared with the E. coli counterpart. The E. coli-derived met-hBMP-2 has shown, by MALDI-TOF-MS, a large presence of the bioactive dimer. A more complex molecular mass (MM) distribution was found for the CHO-derived product, whose exact MM has never been reported because of its variable glycosylation. A method based on RP-HPLC was set up, allowing a quantitative and qualitative hBMP-2 determination even directly on ongoing culture media. Considering that hBMP-2 is highly unstable, presenting moreover an extremely high aggregate value, we believe that these data pave the way to a necessary characterization of this important factor when synthesized by DNA recombinant techniques in different types of hosts. (AU)

FAPESP's process: 16/24724-6 - Expression, purification and characterization of bone morphogenetic protein (BMP-2) for application in composites biomaterials with poly(E-caprolactone)
Grantee:Paolo Bartolini
Support Opportunities: Research Grants - Innovative Research in Small Business - PIPE
FAPESP's process: 15/15446-0 - Expression, purification and characterization of bone morphogenetic protein (BMP-2) for application in composites biomaterials with poly(E-caprolactone)
Grantee:Paolo Bartolini
Support Opportunities: Research Grants - Innovative Research in Small Business - PIPE